Purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated Bim. The final product is generated by affinity chromatography using a Bim-derived peptide that is phosphorylated at serine 65 (serine 69 in the human sequence).
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 17 kDa).
Induces apoptosis. Isoform BimL is more potent than isoform BimEL. Isoform Bim-alpha1, isoform Bim-alpha2 and isoform Bim-alpha3 induce apoptosis, although less potent than the isoforms BimEL, BimL and BimS. Isoform Bim-gamma induces apoptosis.
Isoform BimEL, isoform BimL and isoform BimS are the predominant isoforms and are ubiquitously expressed with a tissue-specific variation. Isoform Bim-gamma is most abundantly expressed in small intestine and colon, and in lower levels in spleen, prostate, testis, heart, liver and kidney.
Belongs to the Bcl-2 family.
The BH3 motif is required for Bcl-2 binding and cytotoxicity.
Mitochondrion and Endomembrane system. Associated with intracytoplasmic membranes.
Western blot - Bim (phospho S65) antibody (ab17935)
Predicted band size : 17 kDa
Lysates prepared from Hek293T cells transfected with rat WT Bim alone (1, 4), cotransfected with rat WT Bim and activated MEKK1 (2, 5) or with
mutant S65A Bim and MEKK1 (3, 6), were resolved by SDS-PAGE on a 14% polyacrylamide gel and transferred to PVDF. Membranes were blocked with 3% Milk-TBST buffer for one hour at room temperature, and incubated with Bim Pan antibody (1, 2, 3) or ab17935 (4, 5, 6) for two hours at room temperature in 3% Milk-TBST buffer. After washing, membranes were incubated with a goat F(ab')2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal™ method.
The data show that the signal is detected only in lysates co-expressing rat WT Bim and active MEKK1. This signal is abolished in cells expressing mutant rat Bim S65A and MEKK1, verifying that the signal is site-phosphospecific.